Small Molecule Screening of Primary Human Acute Myeloid Leukemia Using Co-culture and Multiplexed FACS Analysis
(2022) In Bio-protocol 12(6).- Abstract
Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the differentiation of primary AML cells, thus allowing for mechanistic studies in vitro. Additionally, the co-culture on OP9-M2 stromal is superior in preserving surface marker expression of primary (adult and pediatric) AML cells in comparison to stroma-free culture. Thus, by combining the co-culture with multicolor, high-throughput FACS, we can evaluate the effect of hundreds of small molecules on... (More)
Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the differentiation of primary AML cells, thus allowing for mechanistic studies in vitro. Additionally, the co-culture on OP9-M2 stromal is superior in preserving surface marker expression of primary (adult and pediatric) AML cells in comparison to stroma-free culture. Thus, by combining the co-culture with multicolor, high-throughput FACS, we can evaluate the effect of hundreds of small molecules on multi-parametric processes including: cell survival, stemness (leukemic stem cells), and myeloid differentiation on the primary AML cells at a single-cell level. This method streamlines the identification of potential therapeutic agents, but also facilitates combinatorial screening aiming, for instance, at dissecting the regulatory pathways in a patient-specific manner.
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- author
- Baudet, Aurélie LU ; Hultmark, Simon LU ; Ek, Fredrik LU and Magnusson, Mattias LU
- organization
-
- Division of Molecular Medicine and Gene Therapy
- StemTherapy: National Initiative on Stem Cells for Regenerative Therapy
- Stem cell and Cancer stem cell Regulation (research group)
- Chemical Biology and Therapeutics (research group)
- NanoLund: Centre for Nanoscience
- MultiPark: Multidisciplinary research focused on Parkinson´s disease
- LUCC: Lund University Cancer Centre
- publishing date
- 2022-03-20
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Drug, FACS, High throughput, Leukemia, Screen, Stem cell
- in
- Bio-protocol
- volume
- 12
- issue
- 6
- article number
- e4353
- publisher
- Bio-protocol LLC
- external identifiers
-
- pmid:35434186
- scopus:85128129686
- ISSN
- 2331-8325
- DOI
- 10.21769/BioProtoc.4353
- language
- English
- LU publication?
- yes
- id
- f2d9624d-1e72-41bf-bb9c-7a754e353b13
- date added to LUP
- 2022-07-01 15:02:53
- date last changed
- 2024-09-20 03:03:48
@article{f2d9624d-1e72-41bf-bb9c-7a754e353b13, abstract = {{<p>Ex vivo culture of primary acute myeloid leukemia (AML) cells is notoriously difficult due to spontaneous differentiation and cell death, which hinders mechanistic and translational studies. To overcome this bottleneck, we have implemented a co-culture system, where the OP9-M2 stromal cells support the growth, but most notably limit the differentiation of primary AML cells, thus allowing for mechanistic studies in vitro. Additionally, the co-culture on OP9-M2 stromal is superior in preserving surface marker expression of primary (adult and pediatric) AML cells in comparison to stroma-free culture. Thus, by combining the co-culture with multicolor, high-throughput FACS, we can evaluate the effect of hundreds of small molecules on multi-parametric processes including: cell survival, stemness (leukemic stem cells), and myeloid differentiation on the primary AML cells at a single-cell level. This method streamlines the identification of potential therapeutic agents, but also facilitates combinatorial screening aiming, for instance, at dissecting the regulatory pathways in a patient-specific manner.</p>}}, author = {{Baudet, Aurélie and Hultmark, Simon and Ek, Fredrik and Magnusson, Mattias}}, issn = {{2331-8325}}, keywords = {{Drug; FACS; High throughput; Leukemia; Screen; Stem cell}}, language = {{eng}}, month = {{03}}, number = {{6}}, publisher = {{Bio-protocol LLC}}, series = {{Bio-protocol}}, title = {{Small Molecule Screening of Primary Human Acute Myeloid Leukemia Using Co-culture and Multiplexed FACS Analysis}}, url = {{http://dx.doi.org/10.21769/BioProtoc.4353}}, doi = {{10.21769/BioProtoc.4353}}, volume = {{12}}, year = {{2022}}, }